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Recombinant human collagen and medical nanometer fibrous membrane thereof

A human-derived collagen and nanofiber membrane technology, applied in the fields of peptide technology and biomedical materials, can solve problems such as insufficient research, and achieve the effect of improving quality and production efficiency, non-immunogenicity, and good biocompatibility

Active Publication Date: 2018-01-09
南京艾澜德生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the prior art does not fully study this. For example, Chinese patent application CN105963800A discloses a medical stent material, which is prepared by the following steps: first mix and stir calcium nitrate and deionized water until dissolved; Mix and stir sodium silicate and deionized water until dissolved; add polyethylene glycol to calcium nitrate solution, add sodium silicate solution dropwise while stirring, and continue stirring after dropping; filter, wash with absolute ethanol; put in an oven Dry; mix with chitosan, sodium alginate, gelatin, gum arabic, carboxymethyl cellulose, methyl p-hydroxybenzoate, deionized water and stir at high speed; add glacial acetic acid and stir; put in the refrigerator to freeze; put Freeze drying in a freeze dryer; mix and dissolve sodium hydroxide and deionized water; soak the freeze-dried product in sodium hydroxide solution; wash with deionized water until neutral, and dry to obtain

Method used

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  • Recombinant human collagen and medical nanometer fibrous membrane thereof
  • Recombinant human collagen and medical nanometer fibrous membrane thereof
  • Recombinant human collagen and medical nanometer fibrous membrane thereof

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Experimental program
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Effect test

Embodiment 1

[0051] Embodiment 1 Preparation of recombinant human collagen of the present invention

[0052]According to the research and design of the inventor, the recombinant human collagen (the amino acid sequence of which is shown in SEQ ID NO: 1) was entrusted to Shanghai Sangon Bioengineering Co., Ltd. to synthesize its coding gene and clone it into the pPIC9K vector, and electrotransform it into Pichia pastoris Yeast expression strain GS115. After the positive transformants were cultured in BMGY medium for 24 hours, they were inoculated in fermentation medium (recipe: glycerol 40g, K 2 SO 4 18.2g, H 3 PO 4 26.7ml, CaSO 4 .2H 2 O 0.93g, MgSO 4 14.9g, KOH 4.13g, dilute to 1L with water) culture at 29°C, control dissolved oxygen at 22-25%, after 4 hours of culture, add methanol to a final concentration of 0.5% (V / V) to induce expression, at 29 Continue shaking culture at ℃ for 72 hours, during which 100% methanol is supplemented every 24 hours to keep the final concentration...

Embodiment 2

[0054] Embodiment 2 Preparation of nanofibrous membrane of the present invention 1

[0055] Prepare the aqueous solution that contains 2% (V / V) acetic acid and 0.2% (V / V) ethanol, dissolve recombinant human collagen of the present invention, chitosan (Shanghai Blue Season Biotechnology Co., Ltd., Mw .700-800kD) and polyethylene oxide (Aldrich, Mw.600kD), respectively prepared as 5% (W / V, g / mL) collagen solution, 5% (W / V, g / mL) chitosan solution And 5% (W / V, g / mL) polyethylene oxide solution.

[0056] Mix the collagen solution with the polyethylene oxide solution at a volume ratio of 10:1, add carbodiimide to a final concentration of 5 mM, stir at 37°C for 10 minutes, then add the chitosan solution (the volume ratio of it to the collagen solution 1:1), continue to stir overnight at 37°C. Then the reaction solution was placed in a syringe and spun on an electrospinning machine (Yongkang Leye, ET 2535). The spinning conditions were a voltage of 12kV, a speed of 0.3ml / h, a tempe...

Embodiment 3

[0058] Embodiment 3 Preparation of nanofibrous membrane of the present invention 2

[0059] Preparation contains the aqueous solution of 3% (V / V) acetic acid and 0.3% (V / V) ethanol, dissolves recombinant human collagen of the present invention, chitosan (Shanghai Blue Season Biotechnology Co., Ltd., Mw .700-800kD) and polyethylene oxide (Aldrich, Mw.600kD), respectively prepared as 10% (W / V, g / mL) collagen solution, 8% (W / V, g / mL) chitosan solution And 8% (W / V, g / mL) polyethylene oxide solution.

[0060] Mix the collagen solution and polyethylene oxide solution at a volume ratio of 5:1, add carbodiimide to a final concentration of 10 mM, stir at 37°C for 10 minutes, and then add chitosan solution (volume ratio of it to collagen solution 1:1), continue to stir overnight at 37°C. Then the reaction solution is placed in a syringe and spun on an electrospinning machine. The spinning conditions are a voltage of 14kV, a speed of 0.5ml / h, a temperature of 37°C, and a gauze plate re...

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Abstract

The invention provides a recombinant human collagen and a medical nanometer fibrous membrane thereof. Besides, the invention also provides a preparation method for the medical nanometer fibrous membrane. The method comprises the following steps: mixing and reacting recombinant human collagen, polyethylene oxide and carbodiimide with chitosan and then spinning.

Description

technical field [0001] The invention belongs to the fields of polypeptide technology and biomedical materials, in particular, the invention relates to recombinant human collagen and chitosan-based medical nanofiber membranes. Background technique [0002] The skin is the largest tissue in the human body and plays an important role in regulating moisture, temperature and electrolytes, and defending against the invasion of foreign substances. Wound repair for patients with burns, diabetes, obesity and cancer has become a global health problem. There are about 600,000 cases of tinea pedis caused by diabetes every year, with the risk of infection, amputation and even death. At the same time, ulcers caused by aging and obesity are becoming more and more obvious. At this time, the traditional treatment methods appear to be limited, and it is more and more urgent to find more advanced and effective biological materials to treat them. [0003] However, the prior art does not fully...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C12N15/12C12P21/02A61L26/00A61L27/24A61L27/20A61L27/18D04H1/728D04H1/4382D01D5/00A61K47/10A61K47/36A61K47/42
Inventor 杨树林邓爱鹏康肸
Owner 南京艾澜德生物科技有限公司
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